Team:Hong Kong HKUST/future

ACEB construction: While all fragments of ACEB construct have been obtained, ACEB construct is yet to be completed. The complete construct with EF-1alpha promoter, mitochondrial leader sequence, aceB gene, tag protein and polyadenylation sequence are to be assembled in pSB1C3 backbone.

Test of promoter efficiency: GRP78 and FABP1 promoter efficiency can be measured by fluorescence intensity of green fluorescence protein at different fatty acid concentrations.

Construction of inducible glyoxylate shunt: The inducible promoter is to be fused with two functional genes, aceA and aceB.

Transfection of glyoxylate shunt: The construct of inducible glyoxylate shunt is to be transfected into HepG2 cell to test for its functionality. We plan to place the cell in media of different fatty acid concentrations. The fatty acid uptake rate of inducible glyoxylate shunt will be compared with one of constitutive glyoxylate shunt